OBJECTIVE:To establish a method for simultaneous determination of the content of 14 common metal elements (Mg,Ca,Cr,Mn,Fe,Ni,Cu,Zn,As,Se,Cd,Sb,Ba and Pb)in Ganmao qingre granules. METHODS:Microwave diges-tion-inductively coupled plasma mass spectrometry(ICP-MS)method was conducted. The samples were digested with concentrated nitric acid-hydrogen peroxide(2∶1,V/V),and then ICP-MS was used to determine the mass concentration of 14 metal elements. Li, Sc,Ge,Y,In,Bi and Tb were used as internal standards,and branches and leaves of bush were used as standard substances. RE-SULTS:The correlation coefficient of the standard curves of 14 elements was larger than 0.999 6,detection limits were in the range of 0.002-0.035 μg/L,recoveries were in the range of 80.57%-104.2% and RSDs were in the range of 0.34%-2.71%. CON-CLUSIONS:The method is specific,accurate and sensitive,and suitable for the content determination of 14 common metal ele-ments in Ganmao qingre granules. The total mass concentrations of 6 batches of heavy metals are all meet the requirements of Green Trade Standards of Importing&Exporting Medicinal Plants&Preparations.

OBJECTIVE:To establish the ion chromatography for the mass fractions of 9 inorganic anions (fluoride ion,for-mate radical,chloride ion,nitrite radical,bromide ion,nitrate radical,sulfate radical,oxalate radical and phosphate radical) in Ganmao qingre granules. METHODS:Ion chromatograph was used. The column was Ion Pac AS11-HC with the mobile phase of po-tassium hydroxide solution by gradient elution(concentration:30 mmol/L,time:15 min),the column temperature was room tem-perature,the volume was 25 μl. RESULTS:The 9 inorganic anions were basically achieved separation and had good linear relation-ship with r of 0.999 2-0.999 9,respectively;RSDs of precision,stability and reproducibility tests were less than 3.50%. Average re-covery was 92.47%-103.51%(RSD=0.42%-3.41%,n=6),respectively. CONCLUSIONS:The established method is fast,accu-rate,sensitive and repeatable and can be used for the determination of 9 inorganic anions in Ganmao qingre granules.

Objective To investigate post-traumatic stress disorder (PTSD) and life quality (QOL) of children and adolescents with cancer,and analyze the correlation between PTSD and QOL and study whether PTSD would influence QOL.Methods We investigated PTSD by using the PTSD-SS,and investigated QOL by using the EORTC QLQ-C30 V3,and the data was performed by SPSS 17.0.Results The PTSD-SS total score and the scores of EORTC QLQ-C30 of function field and of whole life quality field were negatively correlated,and the PTSD-SS total score and the scores in symptom field and six single measurement of EORTC QLQ-C30 were positively correlated.Conclusions PTSD of children and adolescents with cancer was closely related with the poor functional status and the high level of symptoms problem.PTSD has affected QOL.The higher level the PTSD is,the lower level of QOL will be.

Objective:To analyze cytokine pattern of systemic onset juvenile idiopathic arthritis (SOJIA) combined with macrophage activation syndrome (MAS) in children, and study the early diagnostic value in MAS.Methods:The clinical data of 157 children with SOJIA from January 2013 to March 2018 in Taizhou Hospital of Zhejiang Province were retrospectively analyzed. Among them, SOJIA combined with MAS was in 15 cases (SOJIA combined with MAS group), and simple SOJIA was in 142 cases (simple SOJIA group). The peripheral blood levels of interleukin (IL)-2, IL-4, IL-6, tumor necrosis factor (TNF)-α and interferon (IFN)-γ were measured by flow cytometry cytometric beads array. The characteristics of cytokine pattern was analyzed.Results:The IL-10 and IFN-γ in SOJIA combined with MAS group were significantly higher than those in simple SOJIA group: 40.5 (7.9, 236.9) ng/L vs. 4.1 (2.0, 98.7) ng/L and 55.8 (18.5, 500.0) ng/L vs. 4.4 (1.4, 30.1) ng/L, and there were statistical differences ( P<0.01); there were no statistical difference in IL-2, IL-4, IL-6 and TNF-α between 2 groups (P>0.05). Pearson correlation analysis result showed that IL-10 was positive correlated with IFN-γ in SOJIA children with MAS ( r = 0.638, P = 0.011). Receiver operating characteristic curve analysis result showed that the area under the curve (AUC) of IFN-γ in predicting MAS was 0.991, 95% CI 0.974 to 1.000, the optimal critical value was 18.45 ng/L, the sensitivity was 92.5%, and the specificity was 95.1%; the AUC of IL-10 in predicting MAS was 0.944 (95% CI 0.893 to 0.996), the optimal critical value was 7.75 ng/L, the sensitivity was 91.7%, and the specificity was 81.7%. Conclusions:The significant increased IL-10 and IFN-γ is helpful for the early diagnosis MAS in children with SOJIA.

Mycobacterium (M.) vaccae is a fast-growing species of saprophytic bacteria that is widely distributed. To understand the host immune responses induced by M. vaccae isolated from bovine submaxillary lymph nodes, C57BL/6 mice were infected with reference strain M. vaccae Bacillus Calmette-Guérin (BCG) and isolated M. vaccae using intraperitoneal injections. Comparison of the bacterial replication and organ pathology between M. vaccae and M. vaccae BCG revealed that M. vaccae was more malignant than M. vaccae in mice. We also demonstrated that serum from the M. vaccae-infected mice contained a higher expression level of gamma-interferon (IFN-γ), tumor necrosis factor alpha, monocyte chemoattractant protein-1, interleukin (IL)-4, IL-12, IL-10 and transforming growth factor beta than did the other groups, especially after week 4. Furthermore, when the numbers of CD3⁺CD4⁺IFN-γ⁺ and CD3⁺CD4⁺IL4⁺ cells in the infected mice were observed by flow cytometry, we found that a powerful T helper 1 (Th1) response was induced by M. vaccae infection, which was associated with the emergence of CD3⁺CD4⁺IFN-γ⁺ cells. However, the Th1 response declined over time, which was associated with appearance of the CD4⁺CD25⁺FoxP3⁺ and CD4⁺CD25⁺CD152⁺Treg cell reaction. In addition, a strong Th2 response was found. Finally, we found that M. vaccae infection increased the production of type I IFNs, which was associated with a reduced Th1 response.
Animals ; Bacillus ; Bacteria ; Chemokine CCL2 ; Flow Cytometry ; Injections, Intraperitoneal ; Interferon-gamma ; Interleukin-10 ; Interleukin-12 ; Interleukins ; Lymph Nodes ; Mice* ; Mycobacterium bovis ; Mycobacterium* ; Pathology ; Transforming Growth Factor beta ; Tumor Necrosis Factor-alpha

Objective:To investigate the prevalence and molecular characteristics of Escherichia coli ( E. coli) producing a novel Shiga toxin 2k subtype in goat in Lanling county, Shandong province. Methods:In November 2019, 512 goat fecal samples were collected from different households in Lanling county, Shandong province. After enriched with EC broth, stx-positive samples were detected by PCR and inoculated in CHROMagar? ECC agar and CHROMagar? STEC agar. The whole genomes of stx-positive strains were sequenced. Based on the genomic senquences, the stx subtype, serotype, multi-locus sequence type and virulence genes of each strain were analyzed. Results:Eighty-six strains of Shiga toxin-producing E. coli (STEC) were isolated from 512 goat fecal samples. Five stx subtypes were identified and 37 strains were positive for stx2k. The 86 STEC strains belonged to 20 O∶H serotypes and 18 different sequence types (STs). Conclusions:STEC strains circulating in goats in Lanling county, Shandong province were heterogeneous in stx subtypes, serotypes and virulence gene profiles, and a certain proportion of strains producing a novel Shiga toxin 2k subtype were detected.

Objective:To observe the expression changes of microRNA(miR)-122 in liver tissue of rats infected with Clonorchis sinensis and its correlation with expression level of inflammatory cytokines. Methods:Totally 24 SPF grade Wistar male rats were selected and randomly divided into a control group (100 μl physiological saline gavage), a 4-week infection group (100 Clonorchis sinensis metacercariae gavage), and an 8-week infection group (100 Clonorchis sinensis metacercariae gavage) based on body weight (100-120 g) using a random number table method, with 8 rats in each group. Starting from the third week of infection, rat feces were collected and directly smeared with physiological saline for identification of Wistar rat animal models infected with Clonorchis sinensis. After 4 and 8 weeks of infection, the rats in the 4- and 8-week infection groups were euthanized, while 4 rats in the control group were euthanized, respectively. The heart blood and left lobe liver tissue and serum samples were collected from each group of rats. Using hematoxylin-eosin (HE) staining to observe liver pathological damage under the light microscope, real-time fluorescence quantitative PCR to detect the expression level of miR-122 in liver tissue, and Luminex 200 liquid suspension chip to detect the expression levels of serum inflammatory cytokines [tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), interleukin-6 (IL-6)]. The correlation between miR-122 and inflammatory cytokines was analyzed using Pearson correlation. Results:Under the light microscope, the morphology of hepatocytes in control group was normal, and no inflammatory cell infiltration was observed. There was inflammatory cells such as lymphocyte, eosinophil and other inflammatory cell infiltration around the portal area in the 4-week infection group. The hepatocytes of the 8-week infected rats were arranged in a disordered manner, with varying degrees of swelling, loose and lightly stained cytoplasm, and some hepatocytes showed watery degeneration; additionally, bile duct dilation and thickening of the bile duct wall were observed in the liver tissue. There were statistically significant differences of liver miR-122 (1.00 ± 0.32, 2.57 ± 0.60, 3.63 ± 1.63), serum TNF-α [(0.14 ± 0.06), (0.43 ± 0.09), (0.61 ± 0.10) ng/ml], and IL-6 expression levels [(0.03 ± 0.01), (1.06 ± 0.24), (1.48 ± 0.33) ng/ml] in control group, 4- and 8-week infection groups ( F = 13.36, 69.99, 82.23, P < 0.001). There was no statistically significant difference in expression level of IL-1β between different groups ( F = 2.15, P = 0.141). The Pearson correlation analysis showed that the expression level of miR-122 was positively correlated with the expression levels of inflammatory cytokines TNF-α and IL-6 ( r = 0.67, 0.80, P < 0.001). Conclusion:Clonorchis sinensis infection can increase the expression of miR-122 in the host liver tissue, and the miR-122 is closely related to the expression levels of inflammatory cytokines TNF-α and IL-6.

Objective To survey the prevalence of greenhouse farmer's lung and related risk factors in part of rural areas of Liaoning Province.Methods Using uniform scheme,procedures and questionnaire,a survey for 5420 farmers(2660 men and 2760 women)with complete data who work inside greenhouses was performed in Shenyang,Xinmin,Chaoyang,and Jinzhou between August 2006 and June 2009.Pulmonary function tests was performed for every active farmer.Results Greenhouse farmer's lung was diagnosed in 308 cases,205 men(66.55%,205/308)and 103 women(33.44%,103/308),a prevalence of 5.7%(308/5420).The prevalence rate of greenhouse farmer's lung in males was significantly higher than that in females(?2=39.93,P0.05).In the 308 cases,the number of patiernts presented with fever chill,cough/sputum,chest tightness/shortness of breath were 180(58.44%),192(62.34%),160(51.95%)respectively,and the number of crepitations,radiological changes,spirometry abnormalities and serum IgE antibodies(+)was 164(53.25%),153(49.68%),147(47.73%)and 136(44.16%)at the time of the study.62.34%(192/308)of patients with greenhouse farmer's lung were mild and 38.66%(116/308)were severe.Conclusion The total prevalence rate of greenhouse farmer's lung in part of rural areas of Liaoning Province was 5.7% and multiple risk factors were associated with the disease.

OBJECTIVETo investigate the serum levels of Th1/Th2 cytokines in children with non-systemic juvenile onset idiopathic arthritis (non-SOJIA).

METHODSClinical data of 41 children with non-SOJIA, including 11 cases of polyarthritis, 10 cases of oligoarthritis and 20 cases of enthesitis related JIA (ERA), admitted in Children's Hospital of Zhejiang University School of Medicine during November 2012 and May 2015 were retrospectively analyzed. Serum levels of Th1/Th2 cytokines including IL-2, IL-4, IL-6, IL-10, TNF-α and IFN-γ were measured by flow cytometry in patients with non-SOJIA, and compared with those in patients with SOJIA (SOJIA group, n=85) and healthy children (control group, n=202); their correlations with erythrocyte sedimentation rate and C reactive protein and CRP were analyzed.

RESULTSCompared with the healthy control group, serum levels of IL-2, IL-6 and IFN-γ were significantly increased in patients with non-SOJIA (2.9 vs. 2.6 pg/mL, 9.9 vs. 6.4 pg/mL, 6.3 vs. 5.1 pg/mL, allP<0.05),while levels of TNF-α and IL-10 were significantly decreased (2.7 vs. 3.9 pg/mL, 2.9 vs. 7.1 pg/mL, both P<0.01). Compared with the SOJIA group, serum levels of IL-6 and IL-10 were significantly decreased in patients with non-SOJIA (9.9 vs. 33.5 pg/mL, 2.9 vs. 4.1 pg/mL, both P<0.01), while levels of IL-4 and IL-10 were significantly increased (3.1 vs. 2.3 pg/mL, 6.3 vs. 4.4 pg/mL, both P<0.05). Serum levels of IL-6 in patients with polyarthritis or ERA were higher than that in patients with oligoarthritis (12.7 and 11.0 vs. 4.2 pg/mL, both P<0.05). A positive correlation of IL-6 or TNF-α level with C reactive protein was observed in patients with ERA.

CONCLUSIONSThe results indicate that Th1/Th2 imbalance and Th1 predominance may exist in children with non-SOJIA; and IL-6 may be involved in the pathogenesis of non-SOJIA children with polyarthritis.

The identification of clinical questions for clinical practice guidelines of Chinese patent medicine （CPM） is important for subsequent evidence retrieval， evaluation of evidence quality， formation of recommendations. This paper described a methodological proposal for the identification of clinical questions for CPM guidelines to highlight the characteristics of Chinese patent medicine and reflect its effect in specific stage of the disease. Considering four aspects， namely， the drug of Chinese patent medicine （D）， the specific disease stage （S）， comparison （C）， and specific outcome （O）， DSCO framework has been proposed to formulate the clinical questions. Multi-source information through scientific research， policy or standard documents， and clinical data are suggested for collecting clinical questions， and clear selection criteria should be set to finalize the clinical questions to be addressed by the guideline. In addition， the above process needs to be transparently and publicly reported in order to ensure the clarity and completeness of the guidelines.